The study of enzymes usually by biochemical methods involves their quantitative and qualitative isolation in active and/or native structural forms followed by analyses/profiling for catalytic levels, enzymes/isozymic molecular forms to discern functional facets, patterning of proteins and/or their polypeptides and their further analyses and constitutes the subject matter of the term ‘proteomics’.
The subject has been associated with isolation of nearly wholesome of protein complement from cells, tissues, organs or organisms for the follow-up of identification, characterization and quantitation, electrophoretic display of protein and polypeptide profiles, monitoring of catalytic levels of enzymes and isozymes, discerining multiple molecular forms of enzymes etc.
Today, the proteomics covers much of the functional analysis of gene products or functional genomics. Merely DNA sequences in databases fail to decipher the biological function and hence, proteomics has become crucial for understanding modulating biological functions and mechanisms. For this reason, it is a key component of modern biotechnology including genetic manipulation and metabolic engineering.
In fact, the understanding of the biological functions of genes is a more difficult proposition than obtaining just the sequences. From a plant (particularly those with secondary metabolite producing pathways) perspective, the situation is more complex due to far little known about the metabolic networks and catalytic/regulatory proteins involved.
The most critical requirement in the proteomics initiative is the quantitative isolation of cellular protein complement from the test tissue/organism. Although, some general protocols for isolation of proteins under buffered conditions near neutrality of pH could be available in the literature, however, these can not be satisfactorily applied to several tissues (like leaf, petal and sepal) of scented geranium.
In fact, so far there is only singular report on a male fertile genotype of scented geranium concerning aqueous isolation and profiling of proteins under non-denaturing conditions and that pertains to the anthers only (S. Tokumasu, F. Yano and M. Kato, 1977, Japan J. Genet. 52:197-205).
But this prior art is hardly of any relevance for the larger proteomic work as (i) most of the cultivated clones of scented geraniums are male sterile, (ii) the procedure can not be applied to other aerial parts like leaf, petal and sepal, (iii) rather than anthers, other aerial parts of plant are important for the proteomic work from the standpoint of metabolic pathways and production of valuable exotic essential oil for use in perfumery, cosmetic and flavour industries and aromatherapy (E. Gildemeister and Fr. Hoffman, 1959, Die atherischen Ole vol 5, p 350, Akademie Verlag, Berlin).
The scientific explanation for the invention is that the aerial parts like leaf lamina, sepal and petal of scented geranium are highly acidic in nature with cell sap pH of about 3.0 (R. S. Sangwan, B. R. Tyagi and N. S. Sangwan, 2000, Ecological method of phytoremediation of alkaline and chemically degraded soils through scented geranium Pelargonium sp., patent filed, NF399/00, CSIR, New Delhi). The acidity of a two volume aqueous extract of these fresh tissues was found to be more than 5 milliequivalents of sodium hydroxide (R. S. Sangwan, B. R. Tyagi and N. S. Sangwan, 2000, Ecological method of phytoremediation of alkaline and chemically degraded soils through scented geranium Pelargonium sp. The U.S. patent application Ser. No. 09/776,502 for the same is recently allowed.